2021年广东省广州市一株埃可病毒11型菌株的分离与鉴定

Isolation and identification of an Echovirus 11 in Guangzhou,Guangdong province in 2021

目的探索疑似埃可病毒病原体感染标本的分离鉴定,了解埃可病毒11型(ECHO 11)生长特性及进化特征。方法将2021年广东省广州市发热、急性扁桃体炎患儿的咽拭子标本接种人横纹肌肉瘤细胞(RD)进行病毒分离培养通过巢式反转录聚合酶链反应及一步法聚合酶链反应等方法对疑似埃可病毒病原体进行鉴定;扩增分离株VP1基因全长序列,选择全球有代表性流行株,绘制进化树,确定分离株的基因型并研究分离株在细胞中的生长动力学。结果从患者咽拭子标本中分离出ECHO 11毒株,命名为Qi-2023。分离株在RD细胞中能有效增殖,病毒滴度随着复制时间的增加而上升,3~12 h之间病毒增殖最快。进化树结果表明该分离株与D型代表株聚类在一起,VP1基因核苷酸相似性为84.42%~99.08%。且与D5亚型同源性最高,VP1基因核苷酸相似性99.08%。结论从疑似肠道病毒感染患者的咽拭子中成功分离出一株ECHO 11,为D5亚型。这些研究将为ECHO 11感染防控提供实验室数据参考。

Objective To explore the isolation and identification of suspected enterovirus pathogen-infected specimens and understand the growth characteristics and evolutionary characteristics of Echovirus 11(ECHO 11).Methods Enterovirus pathogens were identified through nested reverse transcription-polymerase chain reaction and one-step reverse transcriptionpolymerase chain reaction;Throat swab specimens were collected from children with fever and acute tonsillitis in Guangdong,GuangZhou in 2021 to isolate and culture the virus in human malignant embryonic rhabdomyosarcoma cells(RD),thereby studying the growth dynamics of the isolate in cells;According to the amplified full-length sequence of the VP1 gene of the isolate was amplified,a couple of globally representative epidemic strains were selected to draw an evolutionary tree.Based on the previous works,the genotype of the isolate was determined.Results The ECHO 11 strain was isolated from the patient’s throat swab specimen and named Qi-2023.The isolate can effectively proliferate in RD cells,with the virus titer increasing over time.The fastest increase occurred between 3 and 12 hours.The evolutionary tree analysis revealed that this isolate clustered with the D-type representative strain,and the nucleotide similarity of the VP1 gene ranged from 84.42%to 99.08%.The isolate showed the highest homology with the D5 subtype,with 99.08%nucleotide similarity in the VP1 gene.Conclusion A strain of ECHO 11,subtype D5,was successfully isolated from the throat swabs of patients with suspected enterovirus infection.These studies provide valuable laboratory data for the prevention and control of ECHO 11 infection.